In:
Annals of the New York Academy of Sciences, Wiley, Vol. 1257, No. 1 ( 2012-06), p. 67-76
Abstract:
The tight junction protein ZO‐1 (zonula occludens protein 1) has recruiting/scaffolding functions in the junctional complex of epithelial and endothelial cells. Homodimerization was proposed to be crucial for ZO‐1 function. Here, we investigated the ability of ZO‐1 domains to mediate self‐interaction in living cells. We expressed ZO‐1 truncation mutants as fusions with derivatives of green fluorescent protein in tight junction–free HEK‐293 cells and determined self‐association by means of fluorescence resonance energy transfer measurements using live‐cell imaging. We show that both an SH3‐hinge‐GuK fusion protein and the PDZ2 domain self‐associate in our test system. The recombinant PDZ2 domain also binds to ZO‐1 and ZO‐2 in tight junction–forming HT29/B6 cell lysates, as demonstrated by coprecipitation. Both interaction types are of relevance for the function of ZO‐1 in the regulation of the junctional complex in polar cells.
Type of Medium:
Online Resource
ISSN:
0077-8923
,
1749-6632
DOI:
10.1111/nyas.2012.1257.issue-1
DOI:
10.1111/j.1749-6632.2012.06571.x
Language:
English
Publisher:
Wiley
Publication Date:
2012
detail.hit.zdb_id:
2834079-6
detail.hit.zdb_id:
211003-9
detail.hit.zdb_id:
2071584-5
SSG:
11
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